The Involvement of miR-29b-3p in Arterial Calcification by Targeting Matrix Metalloproteinase-2

Abstract

Vascular calcification is a risk predictor and common pathological change in cardiovascular diseases that are associated with elastin degradation and phenotypic transformation of vascular smooth muscle cells via gelatinase matrix metalloproteinase-2 (MMP2). However, the mechanisms involved in this process remain unclear. In this study, we investigated the relationships between miR-29b-3p and MMP2, to confirm miR-29b-3p-mediated MMP2 expression at the posttranscriptional level in arterial calcification. In male Sprague Dawley rats, arterial calcification was induced by subcutaneous injection of a toxic dose of cholecalciferol. In vivo, the quantitative real-time polymerase chain reaction (qRT-PCR) showed that MMP2 expression was upregulated in calcified arterial tissues, and miR-29b-3p expression was downregulated. There was a negative correlation between MMP2 mRNA expression and miR-29b-3p levels (P = 0.0014, R2 = 0.481). Western blotting showed that MMP2 expression was significantly increased in rats treated with cholecalciferol. In vitro, overexpression of miR-29b-3p led to decreased MMP2 expression in rat vascular smooth muscle cells, while downregulation of miR-29b-3p expression led to increased MMP2 expression. Moreover, the luciferase reporter assay confirmed that MMP2 is the direct target of miR-29b-3p. Together, our results demonstrated that a role of miR-29b-3p in vascular calcification involves targeting MMP2.

The miR-29b/Matrix Metalloproteinase 2 Axis Regulates Transdifferentiation and Calcification of Vascular Smooth Muscle Cells in a Calcified Environment

“…Furthermore, MMP2 is a known target of the miR-29 family, members of which directly inhibit MMP2 expression and suppress cancer cell adhesion, invasion and metastasis [17][18][19][20]. We recently reported that miR-29b-3p regulates MMP2 expression in VC in rats and confirmed MMP2 as a target gene of miR-29b-3p [21]. Therefore, herein we test whether miR-29b-p regulates transdifferentiation and calcification of VSMCs in a calcified environment by affecting MMP2 expression in vitro.…”

The Epigenetic Landscape of Vascular Calcification: An Integrative Perspective

“…Among them, 15 (40.5%) have been confirmed or are suspected to promote or aggravate VC in different scenarios, while 22 (59.5%) protect against VC formation or progression. Direct gene targets have been identified for 14 miRNAs with negative VC regulatory capacity, including miR-29a/29b [14], miR-29b-3p [34], miR-30b [16,47], miR-30c [16], miR-30e [25], miR-34b/34c [29,64], miR-125b [13,22] miR-133a [19], miR-135a [30], miR-182 [42], miR-204 [15,41,50], and miR-205 [21], while few direct targets have been identified for miRNAs with VC enhancement ability (miR-34a [39], miR-128-3p [48], and miR-135a-3p [17]). In the following sections, we describe the clinical features of each VC-regulating miRNA based on their propensity for positive or negative vascular influences.…”

“…The MiR-29 family contain three members, namely miR-29a, 29b, and 29c; these members exhibit immunoregulatory ability, affect muscle cell physiology, and modulate cell survival [98]. Experiments based on rat, human VSMCs, and calcified arteries from rat showed that miR-29a/29b/29c could ameliorate [14,34,44] or aggravate [26,32] calcification progression (Table 1). We believe that this controversial finding can be interpreted from the following perspective; first, those with the direct targets of miR-29 family members uncovered [14,34] all concluded that these miRNAs were negative VC regulators.…”

“…Experiments based on rat, human VSMCs, and calcified arteries from rat showed that miR-29a/29b/29c could ameliorate [14,34,44] or aggravate [26,32] calcification progression (Table 1). We believe that this controversial finding can be interpreted from the following perspective; first, those with the direct targets of miR-29 family members uncovered [14,34] all concluded that these miRNAs were negative VC regulators. Among reports concluding miR-29 family members are pro-calcific, Sudo et al showed that VSMCs with miR-29b over-expression and silencing had nearly 30% and 35% higher and lower calcium deposition than control, respectively, while the miR-29a function was unclear [26].…”

“…Panizo et al showed that silencing miR-29b could lead to less severe calcification but over-expressing miR-29b did not exhibit a corresponding pro-calcific effect [32]. On the other hand, reports that support an anti-calcific role of miR-29 family members uniformly identify a compatible trend of calcification changes upon up-and down-regulation of miR-29a/29b [14] and miR-29b-3p [34]. Consequently, we believe that miR-29 family members more likely ameliorate calcification severity than aggravate calcification in light of current evidence.…”